球菌细胞壁的 MAS NMR 实验:1H 和 CPMAS CryoProbe 增强 13C 检测实验的互补性

IF 2 3区 化学 Q3 BIOCHEMICAL RESEARCH METHODS
Alicia Vallet , Isabel Ayala , Barbara Perrone , Alia Hassan , Jean-Pierre Simorre , Catherine Bougault , Paul Schanda
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引用次数: 0

摘要

细菌细胞壁是千亿吨级的大型交联聚合物,其运动振幅范围很广,既有相当坚硬的部分,也有高度柔韧的部分。魔角旋转 NMR 是获取完整细胞壁原子级信息的有力方法。在此,我们研究了不同同核 13C13C 和异核 1H15N、1H13C 和 15N13C 相关实验的灵敏度和信息含量。我们证明,与室温探针相比,CPMAS CryoProbe 的信噪比提高了约 8 倍,或单位质量灵敏度提高了约 3-4 倍。灵敏度提高后,即使是完整的细菌也能获得高分辨率光谱。此外,我们还比较了在 100 kHz 与 55 kHz 下获得的 1H MAS 实验的分辨率和灵敏度。我们的研究为选择提取细胞壁样品原子级细节的实验提供了有用的提示。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

MAS NMR experiments of corynebacterial cell walls: Complementary 1H- and CPMAS CryoProbe-enhanced 13C-detected experiments

MAS NMR experiments of corynebacterial cell walls: Complementary 1H- and CPMAS CryoProbe-enhanced 13C-detected experiments

Bacterial cell walls are gigadalton-large cross-linked polymers with a wide range of motional amplitudes, including rather rigid as well as highly flexible parts. Magic-angle spinning NMR is a powerful method to obtain atomic-level information about intact cell walls. Here we investigate sensitivity and information content of different homonuclear 13C13C and heteronuclear 1H15N, 1H13C and 15N13C correlation experiments. We demonstrate that a CPMAS CryoProbe yields ca. 8-fold increased signal-to-noise over a room-temperature probe, or a ca. 3–4-fold larger per-mass sensitivity. The increased sensitivity allowed to obtain high-resolution spectra even on intact bacteria. Moreover, we compare resolution and sensitivity of 1H MAS experiments obtained at 100 kHz vs. 55 kHz. Our study provides useful hints for choosing experiments to extract atomic-level details on cell-wall samples.

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来源期刊
CiteScore
3.80
自引率
13.60%
发文量
150
审稿时长
69 days
期刊介绍: The Journal of Magnetic Resonance presents original technical and scientific papers in all aspects of magnetic resonance, including nuclear magnetic resonance spectroscopy (NMR) of solids and liquids, electron spin/paramagnetic resonance (EPR), in vivo magnetic resonance imaging (MRI) and spectroscopy (MRS), nuclear quadrupole resonance (NQR) and magnetic resonance phenomena at nearly zero fields or in combination with optics. The Journal''s main aims include deepening the physical principles underlying all these spectroscopies, publishing significant theoretical and experimental results leading to spectral and spatial progress in these areas, and opening new MR-based applications in chemistry, biology and medicine. The Journal also seeks descriptions of novel apparatuses, new experimental protocols, and new procedures of data analysis and interpretation - including computational and quantum-mechanical methods - capable of advancing MR spectroscopy and imaging.
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